The present disclosure relates to a microscopy system, a microscopy method, and a computer-readable recording medium.
In the field of biology, medicine, and the like, there is a user need that when observing a thick object such as a cell nucleus or a stem cell by using a biological microscope having a focal depth level of several tens of μm, users want to promptly specify a region of interest located in a depth direction (Z direction) along the optical axis of an observation optical system. In response to such a need, there is known a technique with which a plurality of images having different focal planes is acquired by performing sequential imaging while shifting the focal plane of the observation optical system along the optical axis, and based on the plurality of images, an all-in-focus image that is focused at each position in the Z direction is generated. The plurality of images having different focal planes acquired in this way is also collectively called a Z stack image.
Examples of the method of generating an all-in-focus image include a method of restoring, using a blur function, a multi-focus image generated by superimposing Z stack images, and a method of extracting a focal area from each of a plurality of images having different focal planes and performing composition.
For example, WO 2011/158498 discloses a technique with which two images focused on the near-end side and the far-end side of an object and an all-in-focus image generated by imaging while sweeping an image sensor on the far-end side from the near-end side of the object are acquired, the images focused on the near-end side and the far-end side are restored using the all-in-focus image, thereby calculating an amount of blur in a partial area in the image, and as a result, a distance from an optical system to the object is acquired and a distance map is created.